Hyperthermophilic Microorganisms Research Articles

Temperature-dependent iron motion in extremophile rubredoxins – no need for ‘corresponding states’

Extremophile organisms are known that can metabolize at temperatures down to − 25 °C (psychrophiles) and up to 122 °C (hyperthermophiles). Understanding viability under extreme conditions is relevant for human health, biotechnological applications, and our search for life elsewhere in the universe. Information about the stability and dynamics of proteins under environmental extremes is an important factor in this regard. Here we compare the dynamics of small Fe-S proteins – rubredoxins – from psychrophilic and hyperthermophilic microorganisms, using three different nuclear techniques as well as molecular dynamics calculations to quantify motion at the Fe site. The theory of ‘corresponding states’ posits that homologous proteins from different extremophiles have comparable flexibilities at the optimum growth temperatures of their respective organisms. Although ‘corresponding states’ would predict greater flexibility for rubredoxins that operate at low temperatures, we find that from 4 to 300 K, the dynamics of the Fe sites in these homologous proteins are essentially equivalent.

Astrobiological implications of the organic and inorganic cyanide utilization by a novel Antarctic hyperthermophilic Pyrococcus strain.

Organic and inorganic cyanides are widely distributed in nature, yet not much is known about the ability of microorganisms to use these compounds as a source of nitrogen and/or carbon at high temperatures (>80°C). Here we studied the capacity of organic and inorganic cyanides to support growth of an hyperthermophilic Pyrococcus strain isolated from Deception Island, Antarctica. This microorganism was capable of growing with aromatic nitriles, aliphatic nitriles, heterocyclic nitriles, amino aromatic nitriles and inorganic cyanides as nitrogen and/or carbon source. This is the first report of an hyperthermophilic microorganism able to incorporate these compounds in its nitrogen and carbon metabolism. Based on enzymatic activity and genomic information, it is possibly that cells of this Pyrococcus strain growing with nitriles or cyanide, might use the carboxylic acid and/or the ammonia generated through the nitrilase enzymatic activity, as a carbon and/or nitrogen source respectively. This work expands the temperature range at which microorganisms can use organic and inorganic cyanides to growth, having important implications to understand microbial metabolisms that can support life on Earth and the possibility to support life elsewhere.

Recombinant Proteins Combining the Inter-domain Region of Pneumococcal Surface Antigen "A" and Adaptive W-type Polypeptide of Thermotoga Bacteria as a Potential Component Base for the Development of New Diagnostics and Genetically Engineered Subunit Vaccines against Pneumococcal Infection

The problems related to the development of pneumococcal vaccines require combination of traditional solutions and alternative systems optimizing this procedure. Recombinant subunit vaccines have undeniable advantages over inactivated and live-attenuated vaccines: they induce cell-mediated and humoral immunologic responses effectively and with high specificity, but without the risks associated with authentic pathogen processing. However, subunit vaccines require specific adjuvants to enhance the immune response or special fusion partners to improve solubility, expression and optimize subsequent fine purification of the protein of interest. In the framework of this work, a structurally conserved region of the most immunogenic region of the vaccine-valuable surface antigen PspA of Streptococcus pneumoniae was chosen as a model protein, and the adaptive polypeptide CheW from the hyperthermophilic microorganism Thermotoga petrophila was used as a promising fusion protein. Appropriate expression plasmid vectors were designed in silico and constructed in vitro. Efficient E. coli producer strains were obtained and appropriate conditions for heterologous production of chimeric proteins were selected. The fusion partner from T. petrophila positively influenced the properties of the resulting constructs such as thermostability, solubility, and homogeneity. During this work, the optimal pH and temperature ranges of the created proteins were determined, and the principles of low-stage purification were elaborated. We obtained and characterized new proteins, which were not previously found in nature in a similar bioconfiguration. The results indicate that the biotechnologically valuable characteristics of the fusion protein were more expressed when the adaptive CheW protein was combined with the N-terminus of the PspA antigen.

Development of a biosecure carcass composting method using a fermentation system containing hyper-thermophilic microorganisms

Development of a biosecure carcass composting method using a fermentation system containing hyper-thermophilic microorganisms

Taxonomic Diversity of the Microbial Biofilms Collected along the Thermal Streams on Kunashir Island

Hot springs are known as highly adverse extreme environments where thermophilic and hyperthermophilic microorganisms can survive. We describe taxonomic diversity of several microbial biofilms collected along water temperature gradient in hot streams in the aquatic system of the Stolbovskie hot springs on Kunashir Island, Kurils, Russia. The taxonomic composition of the studied microbial communities was assessed by the 16S rRNA gene metabarcoding for bacteria and archaea, and by the 18S rRNA gene metabarcoding for protists. Richness and diversity of bacteria in the geothermal microbial communities decreased with the increase of temperature, while for archaea, the tendency was the opposite. Ciliophora was the most represented taxon of protists. The biofilms of various kinds that we found in a very local area of the geothermal system were different from each other by taxonomic composition, and the level of their taxonomic diversity was significantly influenced by water temperature.

A novel SfaNI-like restriction-modification system in Caldicellulosiruptor extents the genetic engineering toolbox for this genus.

Caldicellulosiruptor is a genus of thermophilic to hyper-thermophilic microorganisms that express and secrete an arsenal of enzymes degrading lignocellulosic biomasses into fermentable sugars. Because of this distinguished feature, strains of Caldicellulosiruptor have been considered as promising candidates for consolidated bioprocessing. Although a few Caldicellulosiruptor strains with industrially relevant characteristics have been isolated to date, it is apparent that further improvement of the strains is essential for industrial application. The earlier identification of the HaeIII-like restriction-modification system in C. bescii strain DSM 6725 has formed the basis for genetic methods with the aim to improve the strain's lignocellulolytic activity and ethanol production. In this study, a novel SfaNI-like restriction-modification system was identified in Caldicellulosiruptor sp. strain BluCon085, consisting of an endonuclease and two methyltransferases that recognize the reverse-complement sequences 5'-GATGC-3' and 5'-GCATC-3'. Methylation of the adenine in both sequences leads to an asymmetric methylation pattern in the genomic DNA of strain BluCon085. Proteins with high percentage of identity to the endonuclease and two methyltransferases were identified in the genomes of C. saccharolyticus strain DSM 8903, C. naganoensis strain DSM 8991, C. changbaiensis strain DSM 26941 and Caldicellulosiruptor sp. strain F32, suggesting that a similar restriction-modification system may be active also in these strains and respective species. We show that methylation of plasmid and linear DNA by the identified methyltransferases, obtained by heterologous expression in Escherichia coli, is sufficient for successful transformation of Caldicellulosiruptor sp. strain DIB 104C. The genetic engineering toolbox developed in this study forms the basis for rational strain improvement of strain BluCon085, a derivative from strain DIB 104C with exceptionally high L-lactic acid production. The toolbox may also work for other species of the genus Caldicellulosiruptor that have so far not been genetically tractable.

High current density via direct electron transfer by hyperthermophilic archaeon, Geoglobus acetivorans, in microbial electrolysis cells operated at 80 °C

Utilization of hyperthermophilic electro-active microorganisms in microbial electrolysis cells (MECs) that are used for hydrogen production from organic wastes offers significant advantages, such as increased reaction rate and enhanced degradation of insoluble materials. However, only a limited number of hyperthermophilic bioelectrochemical systems have been investigated so far. This study is the first to illustrate hydrogen production in hyperthermophilic MECs with a maximum rate of 0.57 ± 0.06 m3 H2/m3d, where an iron reducing archaeon, Geoglobus acetivorans, was used as inoculum. In fact, this is the first study to report that G. acetivorans, as the fourth hyperthermophilic electro-active archaeon. In single chamber MECs operated at 80 °C with a set potential of 0.7 V, a peak current density of 1.53 ± 0.24 A/m2 has been attained and this is the highest record of current produced by pure culture hyperthermophilic microorganisms. Turnover cyclic voltammetry curve illustrated a sigmoidal shape (midpoint of −0.40 V vs. Ag/AgCl), and together with linear relation of scan rate and peak anodic current, proves the biofilm attachment to the anode and its capability of direct electron transfer. Along with simple substrate (acetate), G. acetivorans effectively utilized dark fermentation effluent for hydrogen production in MECs.

Thermostable Cellulases / Xylanases From Thermophilic and Hyperthermophilic Microorganisms: Current Perspective.

The bioconversion of lignocellulose into monosaccharides is critical for ensuring the continual manufacturing of biofuels and value-added bioproducts. Enzymatic degradation, which has a high yield, low energy consumption, and enhanced selectivity, could be the most efficient and environmentally friendly technique for converting complex lignocellulose polymers to fermentable monosaccharides, and it is expected to make cellulases and xylanases the most demanded industrial enzymes. The widespread nature of thermophilic microorganisms allows them to proliferate on a variety of substrates and release substantial quantities of cellulases and xylanases, which makes them a great source of thermostable enzymes. The most significant breakthrough of lignocellulolytic enzymes lies in lignocellulose-deconstruction by enzymatic depolymerization of holocellulose into simple monosaccharides. However, commercially valuable thermostable cellulases and xylanases are challenging to produce in high enough quantities. Thus, the present review aims at giving an overview of the most recent thermostable cellulases and xylanases isolated from thermophilic and hyperthermophilic microbes. The emphasis is on recent advancements in manufacturing these enzymes in other mesophilic host and enhancement of catalytic activity as well as thermostability of thermophilic cellulases and xylanases, using genetic engineering as a promising and efficient technology for its economic production. Additionally, the biotechnological applications of thermostable cellulases and xylanases of thermophiles were also discussed.

Diversity and distribution of thermophilic microorganisms and their applications in biotechnology.

Hot springs ecosystem is the most ancient continuously inhabited ecosystem on earth which harbors diverse thermophilic bacteria and archaea distributed worldwide. Life in extreme environments is very challenging so there is a great potential biological dark matter and their adaptation to harsh environments eventually producing thermostable enzymes which are very vital for the welfare of mankind. There is an enormous need for a new generation of stable enzymes that can endure harsh conditions in industrial processes and can either substitute or complement conventional chemical processes. Here, we review at the variety and distribution of thermophilic microbes, as well as the different thermostable enzymes that help them survive at high temperatures, such as proteases, amylases, lipases, cellulases, pullulanase, xylanases, and DNA polymerases, as well as their special properties, such as high-temperature stability. We have documented the novel isolated thermophilic and hyperthermophilic microorganisms, as well as the discovery of their enzymes, demonstrating their immense potential in the scientific community and in industry.

Isolation and thermo-acclimation of thermophilic bacteria in hyperthermophilic fermentation system.

Hyperthermophilic microorganisms play a key role in the hyper-thermophilic composting (HTC) technique. However, little information is available about the hyperthermophilic microorganisms prevalent in HTC systems, except for the Calditerricola satsumensis, Calditerricola yamamurae, and Thermaerobacter. To obtain effective hyper-thermophilic microorganisms, a continuous thermo-acclimation of the suitable thermophilic microorganisms was demonstrated in this study. Bacillus thermoamylovorans with high-temperature endurance (70°C) were newly isolated from sludge composting, and an adequate slow heating rate (2°C per cycle) was applied to further improve its thermostability. Finally, a strain with a maximum growth temperature of 80°C was obtained. Moreover, structural and hydrophobic changes in cell proteins, the special amino acid content ratio, and the membrane permeability of the thermophilic bacterium after thermo-acclimation were evaluated for improved thermostability. In addition, the acclimated hyperthermophilic bacterium was further inoculated into the HTC system, and an excellent performance with a maximum operating temperature of 82°C was observed.

Discovery of hyperstable carbohydrate-active enzymes through metagenomics of extreme environments.

The enzymes from hyperthermophilic microorganisms populating volcanic sites represent interesting cases of protein adaptation and biotransformations under conditions where conventional enzymes quickly denature. The difficulties in cultivating extremophiles severely limit access to this class of biocatalysts. To circumvent this problem, we embarked on the exploration of the biodiversity of the solfatara Pisciarelli, Agnano (Naples, Italy), to discover hyperthermophilic carbohydrate-active enzymes (CAZymes) and to characterize the entire set of such enzymes in this environment (CAZome). Here, we report the results of the metagenomic analysis of two mud/water pools that greatly differ in both temperature and pH (T=85°C and pH 5.5; T=92°C and pH 1.5, for Pool1 and Pool2, respectively). DNA deep sequencing and following insilico analysis led to 14934 and 17652 complete ORFs in Pool1 and Pool2, respectively. They exclusively belonged to archaeal cells and viruses with great genera variance within the phylum Crenarchaeota, which reflected the difference in temperature and pH of the two Pools. Surprisingly, 30% and 62% of all of the reads obtained from Pool1 and 2, respectively, had no match in nucleotide databanks. Genes associated with carbohydrate metabolism were 15% and 16% of the total in the two Pools, with 278 and 308 putative CAZymes in Pool1 and 2, corresponding to ~2.0% of all ORFs. Biochemical characterization of two CAZymes of a previously unknown archaeon revealed a novel subfamily GH5_19 β-mannanase/β-1,3-glucanase whose hemicellulose specificity correlates with the vegetation surrounding the sampling site, and a novel NAD+ -dependent GH109 with a previously unreported β-N-acetylglucosaminide/β-glucoside specificity. DATABASES: The sequencing reads are available in the NCBI Sequence Read Archive (SRA) database under the accession numbers SRR7545549 (Pool1) and SRR7545550 (Pool2). The sequences of GH5_Pool2 and GH109_Pool2 are available in GenBank database under the accession numbers MK869723 and MK86972, respectively. The environmental data relative to Pool1 and Pool2 (NCBI BioProject PRJNA481947) are available in the Biosamples database under the accession numbers SAMN09692669 (Pool1) and SAMN09692670 (Pool2).

Preparation and Characterization of a New Mutant Homolog of the Chemotaxis Protein CheY from the Anaerobic Hyperthermophilic Microorganism Thermotoga Naphthophila

Preparation and Characterization of a New Mutant Homolog of the Chemotaxis Protein CheY from the Anaerobic Hyperthermophilic Microorganism Thermotoga Naphthophila

Гетерологическая экспрессия и характеристика нового мутантного ДНК-связывающего белка из гипертермофильного микроорганизма Thermotoga naphthophila

This paper presents data on cloning, heterogeneous expression and characterization of recombinant homologs of a DNA-binding protein contained in the Thermotoga naphthophila (TnaDBP, TnaDBP-mut) hyperthermophilic microorganism. Initially, the nucleotide sequence of the original thermostable TnaDBP protein was subjected to codon optimisation in accordance with the structural and operational features of the nucleic acid metabolism system of the Escherichia coli mesophilic bacterium subsequently used as a laboratory strain-producer. Expression vector constructs were designed to ensure efficient production of TnaDBP and TnaDBP-mut proteins in E. coli cells. Optimal conditions for the cultivation of transformed strains for the biosynthesis of the soluble form of the target protein product were selected. A trial aerobic cultivation of the microorganisms was carried out under controlled conditions. Specific features of the growth kine - tics for transformed E. coli BL21 (DE3) [pET-TnaDBP-mut] cells were studied. It shown that, under cultivation in a liquid nutrient medium, the E. coli BL21 (DE3) [pET-TnaDBP-mut] culture producing the mutant DNA-binding protein reaches the stationary growth phase 10 hours after the inducer has been administered. A scheme consisting of a few stages is proposed for purifying the obtained proteins using thermolysis. A preliminary assessment of the solubility and thermal stability of the protein according to its primary amino acid sequence was carried out. Possibilities for the practical application of recombinant variants of the thermostable TnaDBP DNA binding protein are considered. Our results evidently demo nstrate that such proteins, due to their unique physicochemical properties, present great interest from a biotechnological point of view and can be used in various industries as sources of essential L -amino acids for cultures of eukaryotic cells as a basis for enteral nutrition of farm animals, as well as a necessary component base in the development of non-viral vector systems and carrier proteins in the field of biomedicine and fundamental science.

Production of Current by Syntrophy Between Exoelectrogenic and Fermentative Hyperthermophilic Microorganisms in Heterotrophic Biofilm from a Deep-Sea Hydrothermal Chimney.

To study the role of exoelectrogens within the trophic network of deep-sea hydrothermal vents, we performed successive subcultures of a hyperthermophilic community from a hydrothermal chimney sample on a mix of electron donors in a microbial fuel cell system. Electrode (the electron acceptor) was swapped every week to enable fresh development from spent media as inoculum. The MFC at 80°C yielded maximum current production increasing from 159 to 247mAm-2 over the subcultures. The experiments demonstrated direct production of electric current from acetate, pyruvate, and H2 and indirect production from yeast extract and peptone through the production of H2 and acetate from fermentation. The microorganisms found in on-electrode communities were mainly affiliated to exoelectrogenic Archaeoglobales and Thermococcales species, whereas in liquid media, the communities were mainly affiliated to fermentative Bacillales and Thermococcales species. The work shows interactions between fermentative microorganisms degrading complex organic matter into fermentation products that are then used by exoelectrogenic microorganisms oxidizing these reduced compounds while respiring on a conductive support. The results confirmed that with carbon cycling, the syntrophic relations between fermentative microorganisms and exoelectrogens could enable some microbes to survive as biofilm in extremely unstable conditions. Graphical Abstract Schematic representation of cross-feeding between fermentative and exoelectrogenic microbes on the surface of the conductive support. B, Bacillus/Geobacillus spp.; Tc, Thermococcales; Gg, Geoglobus spp.; Py, pyruvate; Ac, acetate.

Preparation and characterization of a new mutant homolog of chemotaxis protein CheY from anaerobic hyperthermophilic microorganism Thermotoga naphthophila

Using genetic engineering methods the expression vectors structures have been designed to produce recombinant proteins TnaCheY and Tna CheY-mut, the homologues of the chemotaxis protein CheY from the hyperthermophilic organism Thermotoga naphthophila in Escherichia coli BL21(DE3) cells. The cultivation conditions of transformed strains were optimized. The influence of episomal expression of the heterologous chemotaxis protein CheY on growth kinetics parameters of the culture of mesophilic bacteria E. coli was studied. The optimal purification flowchart of the obtained proteins using thermolysis is proposed. Using the E. coli BL21(DE3) laboratory strain as an example, the possibility of employment the episomal expression of such proteins to control the cultivation and production time of pharmaceutically and industrially valuable metabolites due to the impact on some stages of the bacterial chemotaxis is experimentally proved.

A thermophilic microorganism from Deception Island, Antarctica with a thermostable glutamate dehydrogenase activity

BackgroundThe Antarctic continent is a source of extreme microorganisms. Millions of years of isolation have produced unique biodiversity with adaptive responses to its extreme environment. Although the Antarctic climate is mainly cold, the presence of several geothermal sites, including thermal springs, fumaroles, hot soils and hydrothermal vents, provides ideal environments for the development of thermophilic and hyperthermophilic microorganisms. Their enzymes, called thermoenzymes, are the focus of interest in both academic and industrial research, mainly due to their high thermal activity and stability. Glutamate dehydrogenase, is an enzyme that plays a key role in the metabolism of carbon and nitrogen catalyzing reversibly the oxidative deamination of glutamate to alpha-ketoglutarate and ammonium. It belongs to the family of oxidoreductases, is widely distributed and it has been highly regarded for use as biosensors, particularly for their specificity and ability to operate in photochemical and electrochemical systems. However, the use of enzymes as biosensors is relatively problematic due to their instability to high temperatures, organic solvents and denaturing agents. The purpose of this study is to present the partial characterization of a thermophilic microorganism isolated from Deception Island, Antarctica, that displays glutamate dehydrogenase activity.ResultsIn this work, we report the isolation of a thermophilic microorganism called PID15 from samples of Deception Island collected during the Antarctic Scientific Expedition ECA 46. This microorganism is a thermophile that grows optimally at 50 °C and pH 8.0. Scanning electron microscopy shows rod cells of 2.0 to 8.0 µm of length. Phylogenetic analysis of 16S rRNA gene revealed that this microorganism is closely related to Bacillus gelatini. This microorganism contains a thermostable glutamate dehydrogenase with optimal activity at pH 8.0 and temperatures for its activity from 37 to 50 °C, range of temperature of interest for biotechnological applications. This glutamate dehydrogenase is a highly thermostable enzyme.ConclusionThis is the first report of a microorganism from Antarctica containing a thermostable glutamate dehydrogenase that maintains its activity in a broad range of temperatures making it of potential interest for biotechnological applications.

Upgrade of wood sugar d-xylose to a value-added nutraceutical by in vitro metabolic engineering

The upgrade of D-xylose, the most abundant pentose, to value-added biochemicals is economically important to next-generation biorefineries. myo-Inositol, as vitamin B8, has a six-carbon carbon-carbon ring. Here we designed an in vitro artificial NAD(P)-free 12-enzyme pathway that can effectively convert the five-carbon xylose to inositol involving xylose phosphorylation, carbon-carbon (C-C) rearrangement, C-C bond circulation, and dephosphorylation. The reaction conditions catalyzed by all thermostable enzymes from hyperthermophilic microorganisms Thermus thermophiles, Thermotoga maritima, and Archaeoglobus fulgidus were optimized in reaction temperature, buffer type and concentration, enzyme composition, Mg2+ concentration, and fed-batch addition of ATP. The 11-enzyme cocktail, whereas a fructose 1,6-bisphosphatase from T. maritima has another function of inositol monophosphatase, converted 20 mM xylose to 16.1 mM inositol with a conversion efficiency of 96.6% at 70 °C. Polyphosphate was found to replace ATP for xylulose phosphorylation due to broad substrate promiscuity of the T. maritima xylulokinase. The Tris-HCl buffer effectively mitigated the Maillard reaction at 70 °C or higher temperature. The co-production of value-added biochemicals, such as inositol, from wood sugar could greatly improve economics of new biorefineries, similar to oil refineries that make value-added plastic precursors to subsidize gasoline/diesel production.

Greigite nanocrystals produced by hyperthermophilic archaea of Thermococcales order.

Interactions between hyperthermophilic archaea and minerals occur in hydrothermal deep-sea vents, one of the most extreme environments for life on Earth. These interactions occur in the internal pores and at surfaces of active hydrothermal chimneys. In this study, we show that, at 85°C, Thermococcales, the predominant hyperthermophilic microorganisms inhabiting hot parts of hydrothermal deep-sea vents, produce greigite nanocrystals (Fe3S4) on extracellular polymeric substances, and that an amorphous iron phosphate acts as a precursor phase. Greigite, although a minor component of chimneys, is a recognized catalyst for CO2 reduction thus implying that Thermococcales may influence the balance of CO2 in hydrothermal ecosystems. We propose that observation of greigite nanocrystals on extracellular polymeric substances could provide a signature of hyperthermophilic life in hydrothermal deep-sea vents.

Is there an active hydrothermal flux from the Orca seamount in the Bransfield Strait, Antarctica?

The rifting zone of Bransfield Strait, Antarctica, is tectonically and geologically unique. It is a back-arc basin that was opened by extensional forces associated to roll-back subduction after cessation of spreading activity of the Phoenix Ridge, and the transtension of the westward ending of Scotia-Antarctica Plate boundary. The Bransfield Rift/Ridge is still active generating volcanism or magma rise to force hydrothermal activity. During the ANT-XXV/4 cruise onboard R/V “Polarstern”, standard CTD and beam transmission measurements were done to determine temperature anomaly and turbidity. Water sampling was performed to determine δ3He and to find thermophilic microorganisms to examine the Orca seamount hydrothermal activity. A temperature anomaly of ~0.08 °C, a pick of turbidity, and high value of δ3He (>10%) were found inside Orca seamount. Results are consistent with a hydrothermal flux coming from the seamount. The report of the first observation of thermophilic and hyperthermophilic microorganisms in cold deep Antarctic waters is part of this study. Inside Orca seamount these microorganisms were found at three different depth levels close to the bottom. We suggest that the fluid migration from the volcano resulted from recent magmatic activity and provided the required elemental nutrients for microbial growth. Besides some thermophiles were found outside the seamount in a small quantity close to the seafloor. These would probably be related to subsidiary structures of the Orca seamount, or were transported by currents from other active volcanic sites as Deception Island. The finding of these thermophilic and hyperthermophilic microorganisms raise questions about the dispersal and their resistance in these extreme environments.

Validation of a Hypothesis: Colonization of Black Smokers by Hyperthermophilic Microorganisms.

Newly erupted black smokers (hydrothermal vent chimneys) are sterile during their formation, but house hyperthermophilic microorganisms in substantial amounts in later stages. No direct experimental data exist by which mechanisms hyperthermophiles colonize newly erupted black smokers, but a scenario was proposed recently how this might happen. Here we combine high temperature light microscopy with electron microscopy to show that two hyperthermophilic Archaea, namely Pyrococcus furiosus and Methanocaldococcus villosus are able to adhere onto authentic black smoker material (BSM). We especially are able to directly observe the adhesion process via video recordings taken at high temperatures. These data validate the hypothesis that hyperthermophiles are transferred by serendipitous water currents to the outside of newly formed black smokers and react within seconds to the there prevailing high temperatures by very fast movements. They scan the surface of the hydrothermal chimneys via a much slower zigzag seek-movement and adhere via their flagella at a suitable place, building up biofilms.